The conversion of a dormant bacterial (Bacillus) spore into a vegetative bacterium by the process of spore germination is a relatively simple differentiating system which is readily amenable to biochemical analysis. A major feature of this differentiation process is extensive and rapid hydrolysis of spore protein and RNA. Degradative enzymes are important in regulating enzyme levels especially in mammalian cell, and in several cases specific proteolytic enzymes have been shown to be involved in regulation of several processes including bacterial sporulation, yeast budding, and sea urchin egg development. Therefore, investigation of the role of degradative enzymes in spore germination may provide insights into the control of differentiation in higher organisms. The objective of this project is the elucidation of the role of proteases and ribonucleases in the germination of bacterial spores -especially with regard to potential control functions. Specific aims are to: 1) Determine the function (if any) played by the unique low molecular weight, basic, dormant spore proteins degraded during spore germination; 2) purify and characterize the protease responsible for degrading these unique proteins; 3) determine the location and state (active or inactive) of this enzyme(s) in the dormant spore; 4) elucidate possible control functions for this enzyme in activating, modifying or destroying pre-existing proteins or enzymes; 5) investigate regulation of this protease(s) itself via synthesis of inhibitors or covalent modification; 6) identify the proteins newly synthesized during germination which are subsequently rapidly degraded; 7) elucidate the reason for the rapid proteolysis of these newly synthesized proteins and identify the enzyme(s) involved; 8) determine the dormant spore RNA degraded during germination; 9) purify and characterize the ribonuclease(s) involved in this degradation; and 10) investigate the regulation and function of this enzyme(s) in vivo. BIBLIOGRAPHIC REFERENCES: Setlow, P. Protein Metabolism during Germination of Bacillus Megaterium Spores. In Spore Research, 1975 (Academic Press) in press (1976). Setlow, P. and W.R. Waites. Identification of Several Unique Low Molecular Weight Basic Proteins in Dormant Spores of Clostridium bifermentans and their Degradation during Spore Germination. J. Bacteriol., in press (1976).